Construction of a Quadrangular Tetramer and a Cage-Like Hexamer from Three-Helix Bundle-Linked Fusion Proteins.

Miyamoto T, Hayashi Y, Yoshida K, Watanabe H, Uchihashi T, Yonezawa K, Shimizu N, Kamikubo H, Hirota S, ACS Synth Biol (2019) Europe PMC

SASDFR4 – HP2042 form from Helicobacter pylori, N-terminal domain of syntaxin-1A from Rattus norvegicus, Trp repressor from Escherichia coli (Domain A (PDB:3MLI)-Linker (PDB:1BR0)-Domain B (PDB:1WRT))

HP2042 form from Helicobacter pylori, N-terminal domain of syntaxin-1A from Rattus norvegicus, Trp repressor from Escherichia coli
MWI(0) 177 kDa
MWexpected 150 kDa
VPorod 379 nm3
log I(s) 3.72×101 3.72×100 3.72×10-1 3.72×10-2
HP2042 form from Helicobacter pylori, N-terminal domain of syntaxin-1A from Rattus norvegicus, Trp repressor from Escherichia coli small angle scattering data  s, nm-1
ln I(s)
HP2042 form from Helicobacter pylori, N-terminal domain of syntaxin-1A from Rattus norvegicus, Trp repressor from Escherichia coli Guinier plot ln 3.72×101 Rg: 5.1 nm 0 (5.1 nm)-2 s2
(sRg)2I(s)/I(0)
HP2042 form from Helicobacter pylori, N-terminal domain of syntaxin-1A from Rattus norvegicus, Trp repressor from Escherichia coli Kratky plot 1.104 0 3 sRg
p(r)
HP2042 form from Helicobacter pylori, N-terminal domain of syntaxin-1A from Rattus norvegicus, Trp repressor from Escherichia coli pair distance distribution function Rg: 5.2 nm 0 Dmax: 20 nm

Data validation


Fits and models


log I(s)
 s, nm-1
HP2042 form from Helicobacter pylori, N-terminal domain of syntaxin-1A from Rattus norvegicus, Trp repressor from Escherichia coli CORAL model

SAXS data from solutions of the fusion protein1 in 20 mM Tris-HCl, 150 mM KCl, pH 8.0 were collected using a Nano-Viewer (Rigaku, Japan) at NAIST (Nara, Japan) equipped with a Pilatus 200K detector at a sample-detector distance of 0.75 m and at a wavelength of λ = 0.15418 nm (I(s) vs s, where s = 4πsinθ/λ and 2θ is the scattering angle). Twenty-five successive 60 second frames were collected at a sample temperature of 20°C. Solute concentrations ranging between 2.3 and 8.3 mg/ml were measured. The data were normalized to the intensity of the direct beam and radially averaged. The scattering from the solvent-blank was subtracted. The scattering profile in this entry represents an extrapolation to infinite dilution.

HP2042 form from Helicobacter pylori, N-terminal domain of syntaxin-1A from Rattus norvegicus, Trp repressor from Escherichia coli (Fusion protein1)
Mol. type   Protein
Organism   Helicobacter pylori, Rattus norvegicus, Escherichia coli
Olig. state   Tetramer
Mon. MW   37.4 kDa
Sequence   FASTA
 
PDB code   3MLI
 
PDB code   1BR0
 
PDB code   1WRT